用两性电解质置换色谱法纯化小鼠甲胎蛋白

Purification of mouse alpha-foetoprotein by ampholyte displacement chromatography.

作者信息

Pagé M, Belles-Isles M

出版信息

Can J Biochem. 1978 Sep;56(9):853-6. doi: 10.1139/o78-131.

Abstract

Mouse alpha-foetoprotein (alphaFP) was isolated from H4 hepatoma tissue using Con A-Sepharose salt gradient ion exchange chromatography and ampholyte displacement chromatography, the latter being a new method for a sharp separation of proteins based on their different isoelectric point. The purity of the alphaFP was demonstrated by (i) the absence of contaminant on sodium dodecyl sulphate polyacrylamide gel electrophetic gels, (ii) Ouchterlony's immunodiffusion against monospecific antimouse alphaFP and the absence of precipitation against a polyvalent antinormal mouse serum, (iii) the production of a monospecific antiserum in a rabbit after injection of the purified antigen, and (iv) immunological unreactivity of the produced antiserum against normal hepatic tissue.

摘要

利用伴刀豆球蛋白A-琼脂糖盐梯度离子交换色谱法和两性电解质置换色谱法从H4肝癌组织中分离出小鼠甲胎蛋白（αFP），后者是一种基于蛋白质不同等电点进行蛋白质锐分离的新方法。αFP的纯度通过以下方式得以证明：（i）十二烷基硫酸钠聚丙烯酰胺凝胶电泳凝胶上无污染物；（ii）针对单特异性抗小鼠αFP的双向免疫扩散以及针对多价抗正常小鼠血清无沉淀反应；（iii）注射纯化抗原后在兔体内产生单特异性抗血清；（iv）所产生的抗血清对正常肝组织无免疫反应性。